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Liquid Handling

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https://www.readbyqxmd.com/read/28323828/liquid-handling-lego-robots-and-experiments-for-stem-education-and-research
#1
Lukas C Gerber, Agnes Calasanz-Kaiser, Luke Hyman, Kateryna Voitiuk, Uday Patil, Ingmar H Riedel-Kruse
Liquid-handling robots have many applications for biotechnology and the life sciences, with increasing impact on everyday life. While playful robotics such as Lego Mindstorms significantly support education initiatives in mechatronics and programming, equivalent connections to the life sciences do not currently exist. To close this gap, we developed Lego-based pipetting robots that reliably handle liquid volumes from 1 ml down to the sub-μl range and that operate on standard laboratory plasticware, such as cuvettes and multiwell plates...
March 2017: PLoS Biology
https://www.readbyqxmd.com/read/28319150/high-throughput-transformation-of-saccharomyces-cerevisiae-using-liquid-handling-robots
#2
Guangbo Liu, Clayton Lanham, J Ross Buchan, Matthew E Kaplan
Saccharomyces cerevisiae (budding yeast) is a powerful eukaryotic model organism ideally suited to high-throughput genetic analyses, which time and again has yielded insights that further our understanding of cell biology processes conserved in humans. Lithium Acetate (LiAc) transformation of yeast with DNA for the purposes of exogenous protein expression (e.g., plasmids) or genome mutation (e.g., gene mutation, deletion, epitope tagging) is a useful and long established method. However, a reliable and optimized high throughput transformation protocol that runs almost no risk of human error has not been described in the literature...
2017: PloS One
https://www.readbyqxmd.com/read/28298176/semi-automated-tip-snip-cloning-of-restriction-fragments-into-and-out-of-plasmid-polylinkers
#3
Ichiro Matsumura
Synthetic biologists rely on semi-synthetic recombinant plasmids, but DNA synthesis is constrained by practical limits on length, accuracy, and sequence composition. Cloned DNA parts can be assembled into longer constructs via subcloning, but conventional methods are labor-intensive. One-pot recombination reactions are more convenient but harder to troubleshoot, and those that depend on PCR to create fragments with compatible ends necessitate re-sequencing. The Tip Snip protocol described here enables the subcloning of an insert from one plasmid polylinker into another without PCR or gel purification steps...
March 1, 2017: BioTechniques
https://www.readbyqxmd.com/read/28286575/chiral-determination-of-3-4-methylenedioxypyrovalerone-enantiomers-in-rat-serum
#4
Michael D Hambuchen, Howard P Hendrickson, S Michael Owens
The emerging stimulant drug of abuse (3,4)-methylenedioxypyrovalerone [(R,S)-MDPV] is self-administered as a racemic mixture by intranasal, iv, oral, and smoking routes. The individual enantiomers are known to have widely different pharmacological effects, with (S)-MDPV showing much greater potency than (R)-MDPV in pharmacological testing. The goal of these studies was to develop and validate an analytical method for quantitation of (R)-MDPV, (S)-MDPV and (R,S)-MDPV in small volumes of rat serum using a chiral separation column and liquid chromatography-mass spectrometry...
January 28, 2017: Analytical Methods: Advancing Methods and Applications
https://www.readbyqxmd.com/read/28284032/technical-and-clinical-validation-of-the-greiner-fc-mix-glycaemia-tube
#5
Eline A E van der Hagen, Marion J Fokkert, Amanda M D Kleefman, Marc H M Thelen, Sjoerd A A van den Berg, Robbert J Slingerland
BACKGROUND: Measurement of adequate glucose concentrations is complicated by in vitro breakdown of glucose due to glycolysis. Unlike the commonly used NaF-EDTA and NaF-oxalate phlebotomy tubes, citrated NaF-EDTA tubes are reported to directly and thereby completely inhibit glycolysis. Recently, Greiner introduced the Vacuette® FC-Mix NaF-EDTA-citrate tube, currently the only NaF-citrate tube without volume-disturbing liquid additions available on the European market. Here we present its potential as alternative for the laborious and therefore unfeasible conditions for glucose sampling as recommended by the World Health Organization (WHO)...
March 11, 2017: Clinical Chemistry and Laboratory Medicine: CCLM
https://www.readbyqxmd.com/read/28271103/a-liquid-thermal-gradient-refractive-index-lens-and-using-it-to-trap-single-living-cell-in-flowing-environments
#6
H L Liu, Y Shi, L Liang, L Li, S S Guo, L Yin, Y Yang
A gradient refractive index (GRIN) lens has a great potential for on-chip imaging and detection systems because of its flat surface with reduced defects. This paper reports a liquid thermal GRIN lens prepared using heat conduction between only one liquid, and uses it as a tunable optical tweezer for single living cell trapping in a flowing environment. This liquid GRIN lens consists of a trapezoidal region in the upper layer which is used to establish a GRIN profile by the heat conduction between three streams of benzyl alcohol with different temperatures, and subsequently a rhombus region in the lower layer with compensation liquids to form a steady square-law parabolic refractive index profile only in transverse direction...
March 8, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28269365/a-silicon-neural-probe-fabricated-using-drie-on-bonded-thin-silicon
#7
Xiao Chuan Ong, Amanda Willard, Mats Forssell, Aryn Gittis, Gary K Fedder
A silicon neural probe fabricated using a deep reactive ion etching based process on 250 μm thin silicon wafers was developed. The fabricated probes replicate the design of soft parylene-C based probes embedded in dissolvable needles and can therefore also be used to test the encapsulation properties of parylene-C in-vivo without introducing additional effects introduced by the dissolvable gel. The process also demonstrates the possibility of performing conventional photolithography on substrates bonded to a handle wafer using a backgrinding liquid wax (BGL7080) as an adhesive...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28266605/in-vivo-microsampling-to-capture-the-elusive-exposome
#8
Vincent Bessonneau, Jennifer Ings, Mark McMaster, Richard Smith, Leslie Bragg, Mark Servos, Janusz Pawliszyn
Loss and/or degradation of small molecules during sampling, sample transportation and storage can adversely impact biological interpretation of metabolomics data. In this study, we performed in vivo sampling using solid-phase microextraction (SPME) in combination with non-targeted liquid chromatography and high-resolution tandem mass spectrometry (LC-MS/MS) to capture the fish tissue exposome using molecular networking analysis, and the results were contrasted with molecular differences obtained with ex vivo SPME sampling...
March 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28259254/effects-of-boiling-duration-in-processing-of-white-paeony-root-on-its-overall-quality-evaluated-by-ultra-high-performance-liquid-chromatography-quadrupole-time-of-flight-mass-spectrometry-based-metabolomics-analysis-and-high-performance-liquid-chromatography
#9
Kong Ming, Jun Xu, Huan-Huan Liu, Jin-Di Xu, Xiu-Yang Li, Min Lu, Chun-Ru Wang, Hu-Biao Chen, Song-Lin Li
Boiling processing is commonly used in post-harvest handling of White Paeony Root (WPR), in order to whiten the herbal materials and preserve the bright color, since such WPR is empirically considered to possess a higher quality. The present study was designed to investigate whether and how the boiling processing affects overall quality of WPR. First, an ultra-high performance liquid chromatography quadrupole/time-of-flight mass spectrometry-based metabolomics approach coupled with multivariate statistical analysis was developed to compare the holistic quality of boiled and un-boiled WPR samples...
January 2017: Chinese Journal of Natural Medicines
https://www.readbyqxmd.com/read/28257572/bead-extractor-assisted-ready-to-use-reagent-system-bears-for-immunoprecipitation-coupled-to-maldi-ms
#10
Huiyan Li, Robert Popp, Michael Chen, Elizabeth M MacNamara, David Juncker, Christoph H Borchers
Quantitative protein assays play an important role in the study of biological functions. Immunoassays and mass spectrometry are two main technologies for quantifying proteins in biological samples. The combination of immunoprecipitation (IP) with MALDI technology delivers high assay sensitivity and specificity, but the sample preparation procedure involves multiple washing and transfer steps. These steps can be performed either manually (requiring significant time and labor) or automatically (requiring the purchase of a complex liquid-handling workstation)...
March 16, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28256822/a-simple-method-for-high-sensitivity-determination-of-cosmogenic-35s-in-snow-and-water-samples-collected-from-remote-regions
#11
Mang Lin, Kun Wang, Shichang Kang, Mark H Thiemens
Cosmogenic 35S is useful in understanding a wide variety of chemical and physical processes in the atmosphere, the hydrosphere and the cryosphere. The 87.4-day half-life and the ubiquity of sulfur in natural environments renders it an ideal tracer of many phenomena. Measurements of 35S in snow and water samples are scarce as existing analytical methods require a large volume of sample (>20 L) due to their high analytical activity background and low counting efficiency. Here, we present a new set of snow/water sample collecting and handling procedures for high-sensitivity determination of cosmogenic 35S using a low-level liquid scintillation spectrometer...
March 3, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28251290/tackling-saponin-diversity-in-marine-animals-by-mass-spectrometry-data-acquisition-and-integration
#12
Corentin Decroo, Emmanuel Colson, Marie Demeyer, Vincent Lemaur, Guillaume Caulier, Igor Eeckhaut, Jérôme Cornil, Patrick Flammang, Pascal Gerbaux
Saponin analysis by mass spectrometry methods is nowadays progressively supplementing other analytical methods such as nuclear magnetic resonance (NMR). Indeed, saponin extracts from plant or marine animals are often constituted by a complex mixture of (slightly) different saponin molecules that requires extensive purification and separation steps to meet the requirement for NMR spectroscopy measurements. Based on its intrinsic features, mass spectrometry represents an inescapable tool to access the structures of saponins within extracts by using LC-MS, MALDI-MS, and tandem mass spectrometry experiments...
March 1, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28227607/a-silicon-neural-probe-fabricated-using-drie-on-bonded-thin-silicon
#13
Xiao Chuan Ong, Amanda Willard, Mats Forssell, Aryn Gittis, Gary K Fedder, Xiao Chuan Ong, Amanda Willard, Mats Forssell, Aryn Gittis, Gary K Fedder, Mats Forssell, Aryn Gittis, Gary K Fedder, Amanda Willard, Xiao Chuan Ong
A silicon neural probe fabricated using a deep reactive ion etching based process on 250 μm thin silicon wafers was developed. The fabricated probes replicate the design of soft parylene-C based probes embedded in dissolvable needles and can therefore also be used to test the encapsulation properties of parylene-C in-vivo without introducing additional effects introduced by the dissolvable gel. The process also demonstrates the possibility of performing conventional photolithography on substrates bonded to a handle wafer using a backgrinding liquid wax (BGL7080) as an adhesive...
August 2016: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
https://www.readbyqxmd.com/read/28222253/flow-cytometry-based-rapid-duplexed-immunoassay-for-fusarium-mycotoxins
#14
Árpád Czéh, Miklós Mézes, Francis Mandy, Zsuzsanna Szőke, György Nagyéri, Noémi Laufer, Balázs Kőszegi, Tamás Koczka, Sándor Kunsági-Máté, György Lustyik
At small food processing facilities, the most frequently used test to determine if grain-derived mycotoxin concentrations are compliant with legal limits is the enzyme-linked immunosorbent assay (ELISA). Each kit is designed to detect one of the six dangerous mycotoxins. With the increasing occurrence of coinfection of grain with multiple-mycotoxins in the field and/or during storage, ELISA is no longer a cost effective best assay option. With ELISA, each species of mycotoxin requires different sample preparation/extraction and a 45 min incubation...
February 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28217772/dielectrowetting-manipulation-for-digital-microfluidics-creating-transporting-splitting-and-merging-of-droplets
#15
Hongyao Geng, Jian Feng, Lisa Marie Stabryla, Sung Kwon Cho
Generating, splitting, transporting, and merging droplets are fundamental and critical unit operations for digital (droplet-based) microfluidics. State-of-the-art digital microfluidics performs such operations commonly using electrowetting-on-dielectric (EWOD) in the typical configuration of two parallel channel plates. This paper presents such operations using dielectrowetting (derived from liquid dielectrophoresis), not EWOD, with an array of interdigitated electrodes. The major and unique feature is that the present droplet manipulations are effective for conductive (water with/without surfactant) and non-conductive (propylene carbonate) fluids...
March 14, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28213251/on-line-monitoring-of-in-vitro-oral-bioaccessibility-tests-as-front-end-to-liquid-chromatography-for-determination-of-chlorogenic-acid-isomers-in-dietary-supplements
#16
Daniel Kremr, David J Cocovi-Solberg, Petra Bajerová, Karel Ventura, Manuel Miró
A novel fully automated in-vitro oral dissolution test assay as a front-end to liquid chromatography has been developed and validated for on-line chemical profiling and monitoring of temporal release profiles of three caffeoylquinic acid (CQA) isomers, namely, 3-CQA,4-CQA and 5-CQA, known as chlorogenic acids, in dietary supplements. Tangential-flow filtration is harnessed as a sample processing approach for on-line handling of CQA containing extracts of hard gelatin capsules and introduction of protein-free samples into the liquid chromatograph...
May 1, 2017: Talanta
https://www.readbyqxmd.com/read/28207828/the-wide-spectrum-high-biocidal-potency-of-bioxy-formulation-when-dissolved-in-water-at-different-concentrations
#17
Dori Dagher, Ken Ungar, Richard Robison, Fadi Dagher
Traditional surface disinfectants that have long been applied in medicine, animal husbandry, manufacturing and institutions are inconvenient at best and dangerous at worst. Moreover, some of these substances have adverse environmental impacts: for example, quaternary ammonium compounds ("quats") are reproductive toxicants in both fish and mammals. Halogens are corrosive both to metals and living tissues, are highly reactive, can be readily neutralized by metals, and react with organic matter to form toxic, persistent by-products such as dioxins and furans...
2017: PloS One
https://www.readbyqxmd.com/read/28190338/toward-streamlined-identification-of-dioxin-like-compounds-in-environmental-samples-through-integration-of-suspension-bioassay
#18
Hongxia Xiao, Markus Brinkmann, Beat Thalmann, Andreas Schiwy, Sigrid Große Brinkhaus, Christine Achten, Kathrin Eichbaum, Carolin Gembé, Thomas-Benjamin Seiler, Henner Hollert
Effect-directed analysis (EDA) is a powerful strategy to identify biologically active compounds in environmental samples. However, in current EDA studies, fractionation and handling procedures are laborious, consist of multiple evaporation steps, and thus bear the risk of contamination and decreased recoveries of the target compounds. The low resulting throughput has been one of the major bottlenecks of EDA. Here, we propose a high-throughput EDA (HT-EDA) work-flow combining reversed phase high-performance liquid chromatography fractionation of samples into 96-well microplates, followed by toxicity assessment in the micro-EROD bioassay with the wild-type rat hepatoma H4IIE cells, and chemical analysis of bioactive fractions...
March 21, 2017: Environmental Science & Technology
https://www.readbyqxmd.com/read/28188518/a-robust-protocol-for-protein-extraction-and-digestion
#19
Michelle Atallah, Mark R Flory, Parag Mallick
Proteins play a key role in all aspects of cellular homeostasis. Proteomics, the large-scale study of proteins, provides in-depth data on protein properties, including abundances and post-translational modification states, and as such provides a rich avenue for the investigation of biological and disease processes. While proteomic tools such as mass spectrometry have enabled exquisitely sensitive sample analysis, sample preparation remains a critical unstandardized variable that can have a significant impact on downstream data readouts...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28176961/blooming-gelatin-an-individual-additive-for-enhancing-nanoapatite-precipitation-physical-properties-and-osteoblastic-responses-of-nanostructured-macroporous-calcium-phosphate-bone-cements
#20
Ziba Orshesh, Saeed Hesaraki, Ali Khanlarkhani
In recent years, there has been a great interest in using natural polymers in the composition of calcium phosphate bone cements to enhance their physical, mechanical, and biological performance. Gelatin is a partially hydrolyzed form of collagen, a natural component of bone matrix. In this study, the effect of blooming gelatin on the nanohydroxyapatite precipitation, physical and mechanical properties, and cellular responses of a calcium phosphate bone cement (CPC) was investigated. Various concentrations of blooming gelatin (2, 5, and 8 wt...
2017: International Journal of Nanomedicine
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